CD146在血管炎患者外周血白细胞表达的意义初探

目的探讨血管炎患者外周血白细胞CD146表达与临床活动性间的关系。方法流式细胞术检测39例活动期系统性血管炎患者[显微镜下多血管炎（MPA）13例,韦格纳肉芽肿（WG）9例,变应性肉芽肿性血管炎（CSS）2例,大动脉炎（TA）9例,白塞病（BD）4例,结节性多动脉炎（PAN）2例]及24例系统性红斑狼疮（SLE）患者外周血白细胞CD146表达,其中18例（MPA5例,WG4例,CSS2例,SLE4例,PAN2例,TA1例）患者经糖皮质激素和环磷酰胺治疗后于病情好转时再次检测。结果（1）与健康者相比,血管炎患者活动期中性粒细胞、淋巴细胞CD146表达增多,尤以中性粒细胞最多,差异均有统计学意义（P〈0．05）。（2）中性粒细胞CD146表达与淋巴细胞、单核细胞CD146表达相关（r值分别为0．66、0．853,P=0．000）,与病程、年龄、血沉、C反应蛋白、抗中性粒细胞胞浆抗体（ANCA）、PR3-ANCA、MPO-ANCA、血肌酐、伯明翰血管炎活动指数（BVAS）、系统性红斑狼疮疾病活动指数（SLEDAI）等无明显相关（r值分别为-0．108、-0．059、-0．073、-0．103、0．012、-0．5、-0．232、0．001、-0．08、0．089,P〉0．5）。（3）18例患者经治疗后好转期中性粒细胞、淋巴细胞CD146表达多数呈逐渐减少的趋势（P〈0．05）。结论CD 146在血管炎患者活动期外周血白细胞尤其是中性粒细胞中表达明显升高,随着糖皮质激素和免疫抑制剂治疗病情好转后呈下降或转阴趋势,其在血管炎发病机制中的意义有待深入研究。     

抗中性粒细胞胞质抗体相关性小血管炎发病机制的研究进展

抗中性粒细胞胞质抗体相关性小血管炎是指以微小动脉、毛细血管、小静脉等小血管管壁的炎症和纤维素样坏死为病理特征的一组系统性疾病。目前发现的原发性小血管炎主要有以下几类：韦格纳肉芽肿病（WG）、显微镜下多血管炎（MPA）、变应性肉芽肿性血管炎（CSS）、节段坏死性新月体性肾炎（NCGN）。而抗中性粒细胞胞浆抗体（ANCA）则是一种以中性粒细胞和单核细胞胞浆成分为靶抗原的自身抗体。     

白细胞介素-8受体A型和B型在强直性脊柱炎中的表达及其意义探讨

目的 检测白细胞介素-8受体A型（CXCR1）和白细胞介素-8受体B型（CXCR2）在强直性脊柱炎（AS）患者外周血中性粒细胞、CD14^＋单核细胞和CD3^＋T细胞上的表达水平，探讨其与AS疾病活动的相关性和可能涉及的AS炎症发病机制。方法 研究对象包括30例活动期AS患者，30例活动期类风湿关节炎（RA）患者和30名健康对照者，应用流式细胞术（FCM）检测CXCR1、CXCR2分别在AS患者、RA患者和健康对照者外周血中性粒细胞、CD14^＋单核细胞和CD3^＋T细胞上平均荧光强度（MFI）的表达水平，并和AS患者的临床BASFI、BASDAI、红细胞沉降率（ESR）、血清C反应蛋白（CRP）等指标进行相关分析。结果 CXCR1在AS患者组外周血CD3^＋T细胞上MFI表达水平（41±24）分别较RA患者组（18±10）和健康对照组（19±7）高（P均〈0．01）。CXCR2在AS患者组外周血CD14^＋单核细胞MFI表达水平（210±54）较健康对照组（300±52）低（P〈0．01），与RA患者组（191±53）比较差异无统计学意义（P〉0．05）；CXCR2在AS患者外周血CD14^＋单核细胞上MFI表达下降与患者毕氏疾病功能指数（BASFI）（r=-0．394，P=0．031）、毕氏AS疾病活动指数（BASDAI）（rs=-0．378，P=0．040）、ESR（rs=-0．465，P=0.010）、CRP（rs=-0．648．P=0．000）存在着负相关关系。结论 CXCR1和CXCR2分别在AS患者外周血CD3^＋T细胞和CD14^＋单核细胞表达异常，提示它们可能参与了AS的发病过程。检测AS患者外周血CD14^＋单核细胞的MFI表达水平可能是评价AS疾病活动性有价值的潜在的生物学标志之一。     

急性冠状动脉综合征患者冠状循环内中性粒细胞髓过氧化物酶的变化

目的研究急性冠状动脉综合征患者体循环和冠状循环内中性粒细胞髓过氧化物酶（MPO）浓度梯度的改变及其临床意义.方法 50例行冠状动脉造影患者分为急性心肌梗死（AMI）组10例、不稳定性心绞痛（UA）组20例、稳定性心绞痛（SA）组10例和对照组10例,同时抽取股静脉、主动脉根部及冠状静脉窦血测定中性粒细胞MPO浓度及高敏C反应蛋白（hs-CRP）水平.结果（1）冠心病各亚组均较对照组的低密度脂蛋白（LDL）水平升高（P均〈0.05）.（2）UA组冠状静脉窦血MPO门内百分数显著低于主动脉根部（P〈0.01）,MPO门内百分数在冠状循环显著低于体循环（P〈0.001）;AMI组冠状静脉窦血MPO平均荧光强度显著低于主动脉根部（P〈0.05）,MPO平均荧光强度在冠状循环显著低于体循环（P〈0.001）;而对照组和SA组股静脉、主动脉根部和冠状静脉窦血中的MPO和hs-CRP含量差异均无统计学意义（P〉0.05）.（3）UA组MPO门内百分数与hs-CRP呈正相关（r=0.78,P〈0.01）;与LDL呈正相关（r=0.52,P〈0.05）;AMI组MPO平均荧光强度与hs-CRP呈负相关（r=-0.80,P〈0.05）;与LDL无相关性（r=0.22,P〉0.05）.结论 MPO是反映冠状循环动脉粥样硬化斑块局部炎症的较好指标;通过MPO途径使LDL成为氧化型LDL,可能是斑块不稳定的机制之一.     

慢性持续期支气管哮喘患者外周血中性粒细胞和单核细胞Toll样受体的表达及意义

目的探讨慢性持续期支气管哮喘（简称哮喘）患者外周血中性粒细胞和CD14＋成熟单核细胞TLR2、TLR4表达的变化及意义。方法入选慢性持续期哮喘患者30例,健康对照组46名,应用流式细胞仪全血双色免疫荧光直标法检测中性粒细胞和单核细胞TLR2和TLR4的表达情况,结果以细胞的TLR相对平均荧光强度（rmfi）和TLR相对阳性细胞百分率（rpcp）表示。结果哮喘组患者中性粒细胞TLR2表达的rmfi（1.48±0.19）和rpcp（M=1.34%,Q1-Q3为0.65%～2.91%）均明显低于健康对照组[分别为1.99±0.53和14.13%（Q1-Q3为2.61%～27.51%）,P值均〈0.001];TLR4表达的rmfi（1.77±0.21）明显低于对照组（1.95±0.26）,（P〈0.01）,rpcp（8.16±5.03）%与对照组（9.58±6.40）%比较,差异无统计学意义（P〉0.05）。哮喘组CD14＋单核细胞中TLR2 rpcp（90.92±8.42）%和TLR4 rpcp（11.92±10.34）%明显高于健康对照组[分别为（78.52±18.79）%和（5.78±3.98）%,P值均〈0.01];TLR2 rmfi（9.91±2.14）及TLR4 rmfi（2.60±0.74）与对照组[分别为（9.73±2.71）和（2.77±0.58）]差异无统计学意义（P值均〉0.05）。结论慢性持续期哮喘患者外周血中性粒细胞TLR2和TLR4的表达较健康人群明显下调,CD14＋单核细胞TLR2及TLR4阳性细胞率显著高于健康人群,中性粒细胞和单核细胞TLRs表达的变化在哮喘病程中可能发挥不同的免疫作用。     

抗中性粒细胞胞浆抗体相关性血管炎的药物治疗进展

原发性小血管炎是一类以小动脉、小静脉、毛细血管等发生血管壁炎症、纤维素样坏死为病理特征的一组自身免疫性疾病。在原发性小血管炎中,部分疾病的发生与抗中性粒细胞胞浆抗体（ANCA）密切相关。因此,称之为ANCA相关性血管炎（AAV）。AAV主要包括韦格纳肉芽肿（WG）、显微镜下多血管炎（MPA）、变应性肉芽肿性血管炎（CSS）和局限于肾脏的血管炎（RLV）。     

老年急性心肌梗死患者中性粒细胞黏附分子的表达和溶栓治疗的影响

目的观察老年急性心肌梗死患者中性粒细胞表面黏附分子L-selectin和CD11b的表达以及溶栓治疗对其影响. 方法用免疫荧光技术和流式细胞术测定30例正常对照组和32例老年急性心肌梗死患者溶栓前后中性粒细胞表面黏附分子L-selectin和CD11b的表达. 结果与对照组相比,急性心肌梗死患者中性粒细胞L-selectin的表达下降47.3%（P＜0.01）,而CD11b的表达升高145.5%（P＜0.01）.溶栓治疗前后中性粒细胞L-selectin和CD11b的表达无明显变化. 结论老年急性心肌梗死患者中性粒细胞L-selectin表达下调,CD11b表达上调,提示中性粒细胞被激活.溶栓治疗对中性粒细胞表面黏附分子的表达无影响.     

冠心病患者PTCA前后外周血CD11b、CD62P表达变化及意义

采用流式细胞仪检测23例健康体检者（对照组）及41例冠心病患者（观察组），经皮冠状动脉腔内成形术（PTCA）术前72、2h、术后30min及24、72h外周血中性粒细胞和单核细胞表面CD11b、CD62P的表达。结果观察组患者外周血中性粒细胞和单核细胞CD11b平均荧光强度（MFI）和CD62P阳性细胞百分比较对照组明显升高。PTCA术后24hCD11b荧光强度和CD62P阳性细胞百分比较术前72、2h明显升高。血小板CD62P与中性粒细胞CD11b呈正相关（r=0．253）。认为PTCA术后中性粒细胞和单核细胞CD11b及血小板CD62P表达上调，且可作为反映PTCA后炎症反应和血栓形成状况的指标。     

乙型肝炎肝硬化患者外周血单个核细胞Toll样受体4的变化

目的探讨乙型肝炎肝硬化患者外周血单个核细胞TLR4的变化及意义。方法用流式细胞仪检测30例健康人、40例乙型肝炎肝硬化和30例慢性HBV携带者外周血单个核细胞表面TLR4的表达,采用ELISA法检测血清IL-6水平。结果乙型肝炎肝硬化患者TLR4及IL-6水平分别为13.5±4.6 MFI和80.5±36.5ng/L,均高于正常人（P〈0.01）;慢性HBV携带者TLR4水平为3.8±1.8MFI,与正常人无显著性差异（P〉0.05）,IL-6水平为45.6±36.8ng/L,明显升高（P〈0.01）;乙型肝炎肝硬化患者TLR4表达与IL-6水平呈正相关（γ=0.768,P〈0.01）,而HBV携带者TLR4表达与IL-6水平无显著性相关（γ=-0.775,P〉0.05）;乙型肝炎肝硬化患者TLR4表达及IL-6水平随着Child分级升高而依次升高。结论 TLR4可能与乙型肝炎肝硬化的发病及进展相关。     

鼻息肉组织中ICAM-1、CD62p的表达及意义

留取32例份鼻息肉组织（首发组19例、复发组13例，嗜酸性粒细胞型25例、中性粒细胞型7例）及11例份正常鼻黏膜组织，采用流式细胞仪测定不同标本中的细胞间黏附分子-1（ICAM-1）和P-选择素（CD62q）。结果显示，鼻息肉组织中ICAM-1和CD62q的阳性表达率均明显高于正常鼻黏膜（P均〈0．01）；原发和复发鼻息肉、嗜酸性粒细胞型和中性粒细胞型鼻息肉组织中的ICAM-1和CD62q的阳性表达率差异无统计学意义（P〉0．05）。认为ICAM-1和CD62q在鼻息肉中高表达，但在鼻息肉分型中不起决定作用。     

Chimeric 7e3 Fab (ReoPro) decreases detectable CD11b on neutrophils from patients undergoing coronary angioplasty

Objectives. The purpose of this study was to monitor the effects of chimeric 7E3 Fab (ReoPro) on leukocyte and platelet activation and interaction during coronary angioplasty.Background. Increased expression of CD11b on monocytes and neutrophils promotes their adhesion to endothelial cells, extracellular matrix and smooth muscle cells. Thrombin-activated platelets adhere via P-selectin to monocytes and neutrophils. These cell interactions may affect the outcome of coronary angioplasty.Methods. During coronary angioplasty, venous blood was obtained for flow cytometric detection of leukocyte CD11b; platelet CD41a, CD61a and CD62P; the percentage of leukocytes with adherent platelets and the intensity of bound platelet fluorescence.Results. Leukocyte CD11b expression increased after angioplasty in control patients (neutrophils 171 ± 25 to 255 ± 31 mean fluorescence intensity [MFI, mean ± SEM], n = 25, p < 0.0001; monocytes 200 ± 40 to 248 ± 36 MFI, n = 17, p < 0.05) and decreased in the patients selected to receive chimeric 7E3 Fab (neutrophils 146 ± 30 to 82 ± 22 MFI, n = 25, p < 0.0001; monocytes 256 ± 53 to 160 ± 38 MFI, n = 17, p < 0.05). Neutrophil CD11b decreased after in vitro incubation of whole blood with chimeric 7E3 Fab (n = 5, p = 0.01), but fMLP-induced increases in CD11b were not prevented. The CD11b expression was unchanged and increased with fMLP stimulation after in vitro incubation of isolated neutrophils with chimeric 7E3 Fab. Direct-labeled chimeric 7E3 Fab was not detected bound to neutrophils in whole blood or isolated cells using flow cytometric techniques. Adhesion of isolated neutrophils to protein-coated glass was not prevented by in vitro incubation with chimeric 7E3 Fab. Platelet activation increased after angioplasty in control patients (CD62P 8.9 ± 0.8 to 12.3 ± 1.2 MFI, n = 25, p < 0.05; CD41a 382 ± 25 to 454 ± 26 MFI, n = 25, p < 0.05, CD61a 436 ± 52 to 529 ± 58 MFI, n = 11, p < 0.05); it did not increase in the patients selected to receive chimeric 7E3 Fab (CD62P 13.2 ± 1.0 to 9.0 ± 0.9 MFI, n = 25, p < 0.05; CD61a 398 ± 32 to 410 ± 38 MFI, n = 7, p = NS). Leukocytes with adherent platelets tended to increase in the control group of patients and decrease after the procedure in patients selected to receive chimeric 7E3 Fab; individual and procedure-related variability were marked.Conclusions. Despite standard aspirin and heparin therapy, leukocyte and platelet activation with platelet adherence to leukocytes occurs after coronary angioplasty. Although chimeric 7E3 Fab does not bind to leukocytes directly, it influences CD11b expression in whole blood. Modulation of platelet and leukocyte activation and interaction by chimeric 7E3 Fab may contribute to an improved outcome after coronary angioplasty.     

过敏性哮喘患者外周血CD4~+T细胞表达CD39和CD73水平变化及其临床意义

目的:初步探讨CD4+T细胞表达的腺苷CD39和CD73在过敏性哮喘患者外周血中的水平变化及与叉头蛋白3(FoxP3)、GATA结合蛋白3(GATA3)和维A酸相关孤儿受体(ROR)-γt mRNA相关性。方法:选择54例对屋尘螨过敏的慢性持续期哮喘患者[间歇状态和轻度持续哮喘组(间歇-轻度组)29例,中重度组25例]和45名健康志愿者,通过荧光实时定量PCR方法检测并比较研究对象外周血CD4+T细胞中CD39、CD73、FoxP3、GATA3和ROR-γt mRNA水平。结果:CD39 mRNA水平正常对照组明显高于间歇-轻度组(P=0.000),间歇-轻度组明显高于中重度组(P=0.001)。CD73 mRNA水平正常对照组明显高于间歇-轻度组(P=0.001),哮喘亚组间差异无统计学意义(P=0.126)。哮喘组CD4+T细胞表达的CD39 mRNA与FoxP3 mRNA水平呈显著正相关性(r=0.607,P=0.000),与GATA3 mRNA水平呈显著负相关性(r=-0.505,P=0.000),与ROR-γt mRNA无显著相关性(r=-0.287,P=0.054)。哮喘组CD4+T细胞表达的CD73 mRNA与FoxP3和ROR-γt mRNA水平无显著相关性(r=0.110,P=0.450;r=-0.227,P=0.138),与GATA3mRNA水平呈显著负相关性(r=-0.286,P=0.040)。结论:CD4+T细胞表达的CD39和CD73在过敏性哮喘免疫反应过程中起到重要作用。     

Standardised assessment of membrane proteinase 3 expression. Analysis in ANCA-associated vasculitis and controls

OBJECTIVES: Increased numbers of neutrophils expressing proteinase 3 on their membrane (mPR3) have been reported in anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitis (AAV) and are suggested to be involved in AAV immunopathogenesis. In most studies, neutrophils were analysed for mPR3 expression without priming with TNFalpha, suggesting that mPR3 expression on neutrophils is dependent on other priming events, such as isolation procedures . These priming events can be variable. Therefore, we analysed mPR3 expression on neutrophils before and after priming with TNFalpha to assess whether standardised assessment of mPR3 expression requires priming. Using neutrophils before and after priming with TNFalpha, we assessed percentages of mPR3(+) neutrophils in patients with AAV and in disease and healthy controls. METHODS: Neutrophils from patients with PR3-AAV and MPO-AAV, systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), and from healthy controls were analysed before and after priming with TNFalpha for mPR3 expression. RESULTS: 42% of all individuals analysed showed minimal expression for mPR3 on all neutrophils before priming with TNFalpha, whereas after priming a clear mPR3(+) subset was observed next to mPR3(-) neutrophils, corresponding to bimodal mPR3 expression. In patients with PR3-AAV or MPO-AAV, the percentage of mPR3(+) neutrophils after priming with TNFalpha was significantly increased (p<0.01 and p<0.05, respectively) compared with healthy controls. Percentages of mPR3(+) PMN were also increased in patients with SLE (p<0.01) but not in RA. CONCLUSION: Standardised assessment of proteinase 3 on the membrane of neutrophils requires priming with TNFalpha. Percentages of mPR3(+) PMN are increased in AAV and SLE, but not in RA.     

Coexpression of CD177 and membrane proteinase 3 on neutrophils in antineutrophil cytoplasmic autoantibody–associated systemic vasculitis: Anti–proteinase 3–mediated neutrophil activation is independent of the role of CD177‐expressing neutrophils

Objective

Wegener's granulomatosis (WG) is strongly associated with antineutrophil cytoplasmic autoantibodies (ANCAs) directed against proteinase 3 (PR3). Recent studies have shown that membrane‐bound PR3 (mPR3) is differentially expressed and colocalizes with CD177/NB1 on circulating neutrophils. We undertook this study to assess the differential expression of CD177 on neutrophils from patients with ANCA‐associated systemic vasculitis (ASV) in comparison with patients with systemic lupus erythematosus (SLE), patients with rheumatoid arthritis (RA), and healthy individuals, and to investigate whether colocalization of mPR3 and CD177 affects anti‐PR3–mediated neutrophil activation.

Methods

Expression of CD177 and mPR3 was analyzed by flow cytometry on isolated neutrophils from patients with ASV (n = 53), those with SLE (n = 30), those with RA (n = 26), and healthy controls (n = 31). Neutrophil activation mediated by anti‐PR3 antibodies was assessed by measuring the oxidative burst with a dihydrorhodamine assay.

Results

Percentages of CD177‐expressing neutrophils were significantly higher in patients with ASV and those with SLE than in healthy controls. In 3 healthy donors, CD177 expression was not detected. After priming with tumor necrosis factor α, neutrophils remained negative for CD177 while mPR3 expression was induced. Neutrophils from CD177‐negative donors or CD177− neutrophils sorted from donors with bimodal expression were susceptible to anti‐PR3–mediated oxidative burst. Variation in the extent of anti‐PR3–mediated neutrophil activation among different donors occurred independent of the percentage of CD177‐expressing neutrophils.

Conclusion

Membrane expression of CD177 on circulating neutrophils is increased in patients with ASV and in those with SLE, but not in RA patients. However, primed neutrophils from CD177‐negative individuals also express mPR3 and are susceptible to anti‐PR3–mediated oxidative burst, suggesting that recruitment of CD177‐independent mPR3 is involved in anti‐PR3–induced neutrophil activation.

     

乙型肝炎肝硬化患者外周血T细胞PD-1和PD-L1表达水平及意义

目的探讨乙型肝炎肝硬化患者外周血T淋巴细胞程序性死亡分子-1(PD-1)及其主要配体PD-L1的表达情况。方法在50例乙型肝炎肝硬化患者和25例健康体检者,使用流式细胞仪检测外周血T细胞PD-1和PD-L1表达;采用荧光定量核酸扩增及测序法检测血清HBV DNA载量。结果对照组和肝硬化组外周血T细胞PD-1阳性表达率分别为11.93±1.23%和33.13±3.38%(P<0.05),PD-L1阳性表达率分别为10.59±1.88%和32.47±2.18%(P<0.05);Child-Pugh A级(30.58±2.99%和32.19±1.44%)、B级(34.61±1.43%和33.46±2.58%)和C级(34.2±2.31%和31.76±2.33%)患者外周血T细胞PD-1和PD-L1表达率无显著性相差(P>0.05);在肝硬化患者,T细胞表面PD-l和PD-Ll表达水平与血清HBV DNA载量呈明显正相关(r2=0.8326和:r2=0.643,P<0.05)。结论肝硬化患者外周血T细胞PD-1和PD-L1表达水平明显上调,且与血清HBV DNA载量呈明显正相关,提示T细胞高表达的PD-1可能通过与其配体PD-L1作用而抑制T细胞免疫应答,并导致病毒感染持续。     

NB1 mediates surface expression of the ANCA antigen proteinase 3 on human neutrophils

Antineutrophil cytoplasmic antibodies (ANCAs) with specificity for proteinase 3 (PR3) are central to a form of ANCA-associated vasculitis. Membrane PR3 (mPR3) is expressed only on a subset of neutrophils. The aim of this study was to determine the mechanism of PR3 surface expression on human neutrophils. Neutrophils were isolated from patients and healthy controls, and hematopoietic stem cells from cord blood served as a model of neutrophil differentiation. Surface expression was analyzed by flow cytometry and confocal microscopy, and proteins were analyzed by Western blot experiments. Neutrophil subsets were separated by magnetic cell sorting. Transfection experiments were carried out in HEK293 and HL60 cell lines. Using neutrophils from healthy donors, patients with vasculitis, and neutrophilic differentiated stem cells we found that mPR3 display was restricted to cells expressing neutrophil glycoprotein NB1, a glycosylphosphatidylinositol (GPI)-linked surface receptor. mPR3 expression was decreased by enzymatic removal of GPI anchors from cell membranes and was absent in a patient with paroxysmal nocturnal hemoglobinuria. PR3 and NB1 coimmunoprecipitated from and colocalized on the neutrophil plasma membrane. Transfection with NB1 resulted in specific PR3 surface binding in different cell types. We conclude that PR3 membrane expression on neutrophils is mediated by the NB1 receptor.     

冠心病患者临床类型与细胞外基质金属蛋白酶诱导因子表达水平的关系

目的：研究冠心病临床类型与细胞外基质金属蛋白酶诱导因子（EMMPRIN）的关系。方法根据临床表现和冠状动脉造影结果,179例患者分成稳定型心绞痛（SAP）35例,不稳定型心绞痛（UAP）76例,急性心肌梗死（AMI）68例,健康对照组30例。流式细胞仪检测患者外周血单个核细胞（PBMCs）表面EMMPRIN的平均荧光强度（MFI）;免疫速率法检测血清高敏C反应蛋白（hs-CRP）浓度,分析冠心病组患者EMMPRIN水平与冠状动脉造影Gensini评分的相关性。结果 PBMCs表面EMMPRIN表达在冠心病临床分型中AMI组、UAP组、SAP组明显高于对照组,差异有统计学意义（P＜0.05）。病变血管累及支数越多,其表达增加越明显,三支病变＞双支病变＞单支病变＞对照组,差异有统计学意义（P＜0.05）。相关性分析显示,PBMCs表面EMMPRIN（MFI）与血清hs-CRP浓度及冠状动脉病变Gensini评分呈正相关。结论 EMMPRIN水平与冠心病临床分型相关。     

L-选择素在系统性红斑狼疮患者外周血CD4~+CD25~+T淋巴细胞的表达及其与Foxp3的相关性

目的了解黏附分子L-选择素(L-selectin,CD62L)在系统性红斑狼疮(systemic lupus erythematosus,SLE)患者外周血CD4+CD25+T淋巴细胞的表达,及其与CD4+CD25+Treg细胞转录因子Foxp3的相关性。方法用流式细胞仪检测68例SLE患者外周血CD4+、CD4+CD25+T淋巴细胞CD62L和Foxp3的表达。根据SLEDAI评分,68例SLE患者中活动组36例,稳定组32例;健康对照组35人。结果活动组SLE患者外周血CD4+CD25+CD62L+T淋巴细胞为(1.71±1.60)%,低于对照组的(5.87±3.03)%(P0.01)和稳定组的(4.91±1.69)%(P0.01),差异具有显著性意义;且与SLEDAI呈负相关(r=-0.695,P=0.000),与补体C3水平呈正相关(r=0.522,P=0.000)。活动组SLE患者外周血CD4+CD25+CD62L+Foxp3+T淋巴细胞为(1.06±0.47)%,低于对照组的(3.17±0.87)%(P0.01)和稳定组的(3.46±1.15)%(P0.01),差异有显著性意义。CD4+T淋巴细胞CD62L的表达与CD4+CD25+T淋巴细胞CD62L和Foxp3的表达呈负相关(r=-0.689,P=0.000;r=-0.568,P=0.000);CD4+CD25+T淋巴细胞CD62L的表达与Foxp3的表达呈正相关(r=0.891,P=0.000)。结论CD62L在SLE患者外周血CD4+CD25+T的低表达及与Foxp3表达密切相关,可能在SLE发病中起着重要作用。     

肝内IFN-γ及其分泌细胞在乙型慢加急性肝衰竭发病机制中的作用

目的通过研究肝内促炎细胞因子IFN-γ的表达及IFN-γ的分泌细胞——T淋巴细胞的数量,以明确二者在乙型慢加急性肝衰竭（acute—on—chronic liver failure,ACLF）发病机制中的作用。方法采用免疫组化方法,分析乙型ACLF患者、慢性乙型肝炎（乙肝）患者及正常对照组肝内IFN-γ的原位表达以及其分泌细胞CD4^＋T淋巴细胞与CD8^＋T淋巴细胞的数量。结果①乙型ACLF患者IFN-γ阳性分泌细胞数均明显高于慢性乙肝患者及正常对照组,差异有统计学意义（P均〈0．001）,慢性乙肝患者IFN-γ阳性分泌细胞数明显高于正常对照组,差异有统计学意义（P〈0．001）;②乙型ACLF患者肝内CD4^＋和CD8^＋T淋巴细胞数量较慢性乙肝及正常对照组明显增加（P均〈0．001）,慢性乙肝患者肝内CD4^＋和CD8^＋T淋巴细胞数量较正常对照组明显增加（P均〈0．001）;③肝内IFN-γ阳性分泌细胞数量与CD4＋T淋巴细胞和CD8^＋T淋巴细胞数量均具有明显的相关性（r=-0．896和0．885,P均〈0．001）。结论乙型ACLF患者肝内CD4^＋T淋巴细胞、CD8^＋T淋巴细胞数量的增加,其分泌的IFN-γ的增加可能参与了ACLF的发病过程。     

Toll样受体2介导的Th17细胞活化在HBV感染中的作用

目的探讨乙型肝炎患者外周血单个核细胞(PBMC)中Toll样受体2(TLR2)与Th17细胞的相关性,为阐述HBV感染诱导炎症应答机制提供理论和实验依据。方法选取2012年7月-2013年7月唐都医院感染科门诊和住院的34例乙型肝炎初治患者,其中24例慢性乙型肝炎和10例急性乙型肝炎;另外选取健康对照者10例,分离PBMC,利用HBV C基因型Envelope区肽段(特异性)或佛波酯联合伊屋诺霉素(非特异性)刺激,流式细胞术检测TLR2表达及Th17细胞百分比。进一步用TLR2的激动剂刺激PBMC,检测Th17细胞变化情况。组间比较采用Kruskal-Wallis H检验。结果在非特异性刺激条件下,Th17细胞在慢性乙型肝炎患者体内的百分比(4.08±1.78)%显著高于急性乙型肝炎患者(1.85±1.28)%及健康对照者(2.09±0.53)%(P=0.000 9、0.000 4),而TLR2+及IL-17A+TLR2+的表达在急、慢性乙型肝炎患者与健康人外周血中差异均无统计学意义(P均0.05)。在特异性刺激条件下Th17及TLR2的表达在慢性乙型肝炎患者体内的表达显著高于急性乙型肝炎组[(5.45±1.61)%vs(3.20±1.13)%;(5.19±3.18)%vs(1.88±1.30)%],差异具有统计学意义(P=0.000 6、0.000 6)。加入TLR2激动剂后急、慢性乙型肝炎患者体内Th17细胞的比例均显著升高,但在急性乙型肝炎患者中,刺激前后差异无统计学意义(P0.05)。结论 TLR2可以直接影响Th17细胞的应答,从而促进乙型肝炎中炎症应答反应。